Pedersen M E, Cohen M, Schotz M C
J Lipid Res. 1983 May;24(5):512-21.
The functional (heparin-releasable) fraction of myocardial lipoprotein lipase (LPL) has been located at the lumen surface of capillary endothelium by means of an indirect immunocytochemical perfusion method for electron microscopy. The primary step immunoreactant was an IgG fraction of goat antiserum directed against LPL from rat heart. The second step antibody, conjugated with horseradish peroxidase, was rabbit IgG directed against goat IgG. Peroxidase reaction product, when present, appeared at the surface an in invaginations of the lumenal plasma membrane of capillary endothelium and also on chylomicrons adherent to that membrane. The highest coverage by such product occurred when the highest heparin-releasable heart LPL activity was attained after fat-feeding of rats. Coverage was low when a low level of heparin-releasable heart LPL activity was induced by carbohydrate-feeding. Coverage was very low in the perfused hearts after heparin-release of functional LPL activity. The positive association between these immunocytochemical results and actual levels of functional LPL activities indicates that functional LPL in the isolated rat heart is at the lumen surface of capillary endothelium.
采用间接免疫细胞化学灌注法进行电子显微镜观察,已将心肌脂蛋白脂肪酶(LPL)的功能性(可被肝素释放的)部分定位于毛细血管内皮的管腔表面。初级免疫反应物是山羊抗大鼠心脏LPL抗血清的IgG部分。与辣根过氧化物酶结合的第二步抗体是兔抗山羊IgG。过氧化物酶反应产物(若存在)出现在毛细血管内皮管腔质膜的表面和内陷处,以及附着于该膜的乳糜微粒上。当大鼠喂食脂肪后达到最高的可被肝素释放的心脏LPL活性时,此类产物的覆盖度最高。当通过喂食碳水化合物诱导出低水平的可被肝素释放的心脏LPL活性时,覆盖度较低。在功能性LPL活性被肝素释放后,灌注心脏中的覆盖度非常低。这些免疫细胞化学结果与功能性LPL活性的实际水平之间的正相关表明,分离的大鼠心脏中的功能性LPL位于毛细血管内皮的管腔表面。